An Optical Configuration of Crossed-Beam Photothermal Lens Spectrometer Operating at High Flow Velocities and Its Application for Cysteine Determination in Human Serum and Saliva.

Abstract

Photothermal lens spectrometry (TLS) is a high sensitive technique for trace determination of nonfluorescent materials. Previous photothermal lens spectrometers suffer from operating limitations at high flow velocities, arising from taking the heated element off the probe beam direction, which results in a decrease in the thermal lens (TL) signal. Herein, we describe an optical configuration of the crossed-beam photothermal lens in transversal flow mode in which the propagating direction of the probe beam and liquid sample flow azimuth are concentric (CBTC). The system consists of a microfluidic cell with a volume of lower than 3 μL. In the current optical configuration, using 1-(2-pyridylazo)-2-naphthol (PAN) in ethanol as a test solution, by increasing the sample flow velocity and without increasing chopping frequency, the reduction in sensitivity is less pronounced. Under a 15 Hz chopping frequency, the optimum sample flow velocity is about 2 cm s-1, which is among the highest reported values achieved to date for photothermal lens spectrometers. Although the system operates at higher flow velocities and lower chopping frequencies compared to the collinear configuration, it provides a comparable analytical limit of detection. This optical configuration has been successfully employed for highly sensitive and selective determination of cysteine in human serum and saliva samples through a competitive complexation reaction with Cu-PAN as a colorimetric probe. The detection limit of this method (9.5 nM) shows a significant enhancement (726-times) in comparison to UV-vis measurements.